title: Improved survival and function of rat cryopreserved islets by coculture with sertoli cells.
abstract: In order to investigate how to improve the function and survival of cryopreserved islets, we cocultured cryopreserved thawed rat islets with rat Sertoli cells. After thawing, the islets were divided into the Sertoli cell coculture group and the control group. Using light and transmission electron microscopes, we examined the morphology of islets and measured their apoptosis index (AI) and insulin release stimulation index (SI). Moreover, we measured apoptosis protein and mRNA by western-blot and reverse transcription polymerase chain reaction and cytokine concentrations in supernatant by ELISA. We examined islet graft survival time in diabetic mice and detected insulin in grafts by immunohistochemistry. We found that the morphology, AI, and SI of the coculture group were all significantly improved. The relative expression levels of cleaved caspase-3 P20, P11, and caspase-7 in the coculture group were lower than those in the control group. Compared with the control group, the expression level of Bax was decreased, but that of Bcl-2 was increased. After transplantation, islet survival in the coculture group was similar to that of fresh islets but longer than that in the control group. These results suggest that coculture with rat Sertoli cells significantly improves the yield and function of rat cryopreserved thawed islets by effectively reducing islet apoptosis.
authors.affiliation: Department of Renal Transplantation, Center of Nephropathy, The First Affiliated Hospital, Medicine College, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: Wujun
authors.initials: W
authors.lastname: Xue
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Tian
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Ding
authors.forename: Puxun
authors.initials: P
authors.lastname: Tian
authors.forename: Xinshun
authors.initials: X
authors.lastname: Feng
authors.forename: Yong
authors.initials: Y
authors.lastname: Song
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Luo
authors.forename: Hongbao
authors.initials: H
authors.lastname: Liu
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Wang
authors.forename: Chenguang
authors.initials: C
authors.lastname: Ding
mesh_terms: D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods*; D015925:Cryopreservation; D003921:Diabetes Mellitus, Experimental / Q000601:surgery; D006085:Graft Survival; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D016381:Islets of Langerhans Transplantation; D008297:Male; D051379:Mice; D008819:Mice, Nude; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000166:cytology*; D051028:bcl-2-Associated X Protein / Q000235:genetics
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; D053148:Caspase 3; D053179:Caspase 7